Anti-replicative recombinant 5S rRNA molecules can modulate the mtDNA heteroplasmy in a glucose-dependent manner.

摘要: Mutations in mitochondrial DNA are an important source of severe and incurable human diseases. The vast majority these mutations heteroplasmic, meaning that mutant wild-type genomes present simultaneously the same cell. Only a very high proportion (heteroplasmy level) leads to pathological consequences. We previously demonstrated targeting small RNAs designed anneal with mtDNA can decrease heteroplasmy level by specific inhibition replication, thus representing potential therapy. have also shown 5S ribosomal RNA, partially imported into mitochondria, be used as vector deliver anti-replicative oligoribonucleotides mitochondria. So far, efficiency cellular expression recombinant rRNA molecules bearing therapeutic insertions remained low. In study, we new versions large deletion which causes KSS syndrome, analyzed their annealing import mitochondria cultured cells. To obtain increased stable expression, created transmitochondrial cybrid cell line site for Flp-recombinase this system recombinase-mediated integration genes coding rRNAs nuclear genome. cells induce shift mutation mtDNA. This was directly dependent on sequence insertion. Quantification copy number transfected revealed absence non-specific effect wild type indicating decreased between is not consequence random repopulation depleted pool genomes. change could modulated growth conditions, namely culturing carbohydrate-free medium, forcing them use oxidative phosphorylation providing selective advantage improved respiration capacities. discuss advantages limitations approach propose further development strategy based RNA